Quantitative analysis of 5-hydroxymethylcytosine (5hmC) has remarkable clinical significance to early cancer diagnosis; however, it is limited by the requirement in current assays for large amounts of starting material and expensive instruments requring expertise. Herein, we present a highly sensitive fluorescence method, termed hmC-TACN, for global 5hmC quantification from several nanogram inputs based on terminal deoxynucleotide transferase (TdT)-assisted formation of fluorescent copper (Cu) nanotags. In this method, 5hmC is labeled with click tags by T4 phage β-glucosyltransferase (β-GT) and cross-linked with a random DNA primer via click chemistry. TdT initiates the template-free extension along the primer at the modified 5hmC site and then generates a long polythymine (T) tail, which can template the production of strongly emitting Cu nanoparticles (CuNPs). Consequently, an intensely fluorescent tag containing numerous CuNPs can be labeled onto the 5hmC site, providing the sensitive quantification of 5hmC with a limit of detection (LOD) as low as 0.021% of total nucleotides (S/N = 3). With only a 5 ng input (∼1000 cells) of genomic DNA, global 5hmC levels were accurately determined in mouse tissues, human cell lines (including normal and cancer cells of breast, lung, and liver), and urines of a bladder cancer patient and healthy control. Moreover, as few as 100 cells can also be distinguished between normal and cancer cells. The hmC-TACN method has great promise of being cost effective and easily mastered, with low-input clinical utility, and even for the microzone analysis of tumor models.
Yiqi Liangxue Jiedu prescription (YLJP), a Chinese medicine that is commonly used to prevent liver cancer and is authorized by a national patent (patent No. ZL202110889980.5) has a therapeutic effect on precancerous lesions; however, the underlying mechanism remains unclear. This study is aimed at determining the clinical therapeutic efficacy of YLJP in patients with precancerous liver lesions and to explore and validate its possible effector mechanism.
Kernel abortion in maize (Zea mays L.) can cause significant reductions in yield. Abortion usually commences early in development and frequently occurs because of stresses such as water deficit or low light. Polyamines have been implicated in development processes of fruits and seeds, but in maize the changes of polyamine levels during kernel growth are still unclear. The objectives of this study were to investigate the roles of polyamines in the early stages of kernel growth and kernel abortion. Two single-cross hybrids with different kernel abortion characteristics were grown in a greenhouse with shade treatment. During early development of normal, naturally aborting, and shade-induced aborting kernels, temporal changes in the concentrations of endogenous putrescine (PUT), spermidine (SPD) and spermine (SPM) and their free, perchloric acid (PCA)-soluble conjugated, and PCA-insoluble bound forms, were measured. Changes in DNA content and the number of endosperm nuclei were also measured. Total polyamine levels in normal kernels rapidly increased 4 to 6 d after pollination (DAP), and peaked during active endosperm cell division. The most abundant polyamines were, in order, PCA-soluble conjugated PUT, PCA-soluble conjugated SPD, and free PUT. Aborting kernels had significantly lower levels of polyamines than normal kernels after 4 DAP. The low polyamine levels were temporally associated with a low number of endosperm nuclei and low DNA content. The results suggest that polyamines may be involved in the regulation of early endosperm development in maize.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)